反刍月形单胞菌乙酸生成缺陷株的构建及相关酶活性分析  

Construction of Selenomonas ruminantium with the Acetic Acid Generation Deficiency and Analysis on the Relative Enzyme Activity

作　　者：苑学[1,2] 孙国权[1,2] 逄晓阳[1,3] 刘国文[1] 王哲[1] 

机构地区：[1]吉林大学农学部畜牧兽医学院,吉林长春130062 [2]内蒙古民族大学动物科学技术学院,内蒙古通辽028000 [3]国家农业深加工产品质量监督检验中心,吉林长春130022

出　　处：《安徽农业科学》2011年第25期15390-15392,15394,共4页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金资助项目(30230260;30600441)

摘　　要：[目的]构建反刍月形单胞菌乙酸生成的缺陷株并分析其发酵特性。[方法]应用转座子标签法,通过转座子供体菌E.coliS17-1/pZJ25∷Tn5对受体菌反刍月形单胞菌进行转座子诱变,采用含卡那霉素和氟乙酸纳的选择性培养基筛选接合子。[结果]共筛选出稳定的对卡那霉素和氟乙酸具有抗性的转座工程菌7株。对反刍月形单胞菌的突变株进行16S rRNA鉴定和Tn5的PCR鉴定,及乙酸激酶(AK)和磷酸乙酰转移酶(PTA)酶比活力分析,确定突变株属于pta基因缺失型氟乙酸抗性菌株。[结论]该研究为进一步研究反刍兽瘤胃微生物乙酸的细胞代谢网络和调控奠定基础。[Objective] The research aimed to the construct the acetic acid generation deficiency strain of Selenomonas ruminantium and analyze the fermentation characteristic.[Method] Based on the transposon tagging method,Selenomonas ruminantium（recipient strain） was carried out the transposon mutagenesis by using the transposon donor strain E.coli S17-1/pZJ25∷Tn5.The zygote was screened by using the selective medium which included kanamycin and sodium fluoroacetate.[Result] Seven transposition engineering strains which had the stable resistance to kanamycin and fluoroethanoic acid were screened.The mutant strain of Selenomonas ruminantium was carried out 16S rRNA identification and Tn5 PCR identification.Moreover,the specific activities of AK and PTA were analyzed.The mutant strain belonged to pta gene deletion and fluoroethanoic acid resistance one.[Conclusion] The research laid the foundation for further studying the acetic acid cellular metabolic network and regulation of rumen microorganism in the ruminant animal.

关 键 词：反刍月形单胞菌 转座子 磷酸乙酰转移酶 乙酸激酶 

分 类 号：S188.4[农业科学—农业基础科学]